Date of Award:

5-1-1972

Document Type:

Thesis

Degree Name:

Master of Science (MS)

Department:

Biology

Department name when degree awarded

Zoology (Physiology)

Committee Chair(s)

LeGrande C. Ellis

Committee

LeGrande C. Ellis

Committee

James Gessaman

Committee

Joseph Street

Abstract

Two radiometric techniques are described for the measurement of monoamine oxidase activity in testicular tissue preparations. These techniques utilize 5-HT-2-14C (serotonin) as the substrate and measure its conversion into 5-HIAA-14C (hydroxy-indole acetic acid). The first technique described uses a substrate concentration of 0.9 µM (0.05 µCi) in an incubation volume of 3.1 ml and 120 mg of teased-tubules. Samples are incubated 1 hour. This assay was found suitable for use with teased-tubule preparations but not for preparations containing interstitual cells. An inhibitor was present in the interstitual cell preparations which at the low substrate concentrations used in the first procedure inhibits the MAO activity of the samples. The advantage of this procedure is that a small substrate concentration can be used. The second procedure described utilizes a substrate concentration of 0.65 mM (0.5 µc) in an incubation volume of 60 µl and 2 mg of homogenized testicular tissue. Samples are incubated 30 minutes. This assay was found suitable for homogenized teased-tubules and homogenized whole testicular tissue preparations. Optimal substrate concentration using this procedure is 0.75 mM. The monoamine oxidase activity of the whole testicular tissue preparations was found to have the same MAO activity as the teased-tubule preparations. This method was found to be a rapid, simple, reliable method for the measurement of testicular monoamine oxidase activity.

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